Progress 07/01/22 to 10/31/23

Outputs
Target Audience:Introduction of Technology to Swine Veterinarians and US Pork Producers Introduction The National Pork Producers Council (NPPC) reports that there are over 60,000 pork producers in the US, which generate annual revenues of over $20 billion. Additional sectors tied to pig production, including transportation, animal care, and consumer-related purchases, bring the entire industry to an estimated $122 billion. Our company's target market is aimed at two substantial aspects of this industry: the castration of male pigs and the grow-out business. According to the NPPC, the United States contributed approximately 13.37% to the world's pork production in 2023, collectively producing over 132.95 million pigs each year in the US alone. Assuming 50% of pigs taken to market are males, almost 66 million pig castrations would have been performed in 2021 in the US. Initially, our focus will be on US pork producers before we expand to producers globally. To better understand the industry and its needs, we've conducted several conversations with swine veterinarians and pork producers, exploring the viability of our new product, EpiFix™. Throughout this process, we introduced our technology to Swine Veterinarians and US pork producers, providing an explanation of the scientific background, and gathering their feedback. Our effort is to position EpiFix™ as an alternative to both surgical castration and Improvest®, the only FDA-approved product currently being used to delay the onset of puberty in male pigs. Insights from a Swine Veterinarian The veterinarian highlighted the following key elements: Improvest® offers performance benefits such as improved feed conversion rate (FCR) and growth rate by allowing pigs to develop as intact boars for a longer period than castrated barrows, up until 3-5 weeks before slaughter. Our new product, EpiFix™, may not offer these same performance benefits as pigs will grow with lower levels of endogenous androgens. Resistance to adopting the use of Improvest® comes primarily from retailers due to concerns associated with genetically modified organisms (GMO). Note that meat from Improvest®-treated pigs does not fall under the category of GMOs, but consumer perception is not influenced by this fact. Despite the resistance, the benefits of Improvest®, including a net gain of $4-5 per pig after accounting for the cost of administration, outweigh the inconveniences of administering two doses. Labor and safety costs associated with castration are minimal, with occasional injuries from blade use not being a significant concern. Views from a Large-Scale Hog Producer The producer discussed several industry practices, public perceptions, and the potential of EpiFix™: The producer expressed willingness to try EpiFix™, though they believe customer acceptance is unlikely due to some unfounded fears. Castration is typically performed within 3-5 days of birth in the industry. Improvest® has high brand awareness and offers significant production advantages, estimated at $2.5-$4.5 more per pig. The producer sees a single early shot as a major management advantage, especially since it would be given at the traditional time reserved for castration. Public perception of Improvest® is mixed. Despite evidence of its benefits, the public does not seem to perceive these advantages. Concerns exist about the human safety of handling EpiFix™, similar to those associated with Improvest®. The producer's market currently does not use Improvest® due to customer policies, notably Clemens Foods. Animal welfare and concerns about the use of hormonal treatments are weighed against surgical castration and EpiFix™, respectively, with hormone use often considered more important. Castration costs range from $0.15-$0.20 per pig, but poor technique or the pig's genetics can result in significant animal losses (up to 50-75 pigs per week in a farm that weans 5,000 pigs per week). The producer is interested in the human safety, administration, and potential meat residue aspects of EpiFix™ compared to Improvest®. Insights from the communication with the market Understanding the views of industry experts and large-scale producers is critical for the potential adoption and success of new products like EpiFix™. While concerns exist, there is a recognition of the benefits and potential improvements these alternatives can bring to swine production. However, customer acceptance and market dynamics will ultimately play a significant role in their widespread adoption. Despite the proven performance of Improvest®, it has not been widely adopted in the US market. There are two primary reasons for this. First, there is a lingering negative perception of surgical castration of pigs among the public, and there is no legal basis to prohibit it. Second, consumers have vague concerns about new technologies. From the consumer's perspective, surgical castration is seen as a safer method compared to administering drugs. For consumers, the perceived safety of the meat takes precedence over concerns about animal pain or required labor associated with surgical castration. Considering these factors, our approach to educating the public and marketing our technology includes the following: 1) Utilize the performance comparison results with Improvest® for business purposes but minimize or avoid its use in marketing. 2) Emphasize that our technology is potentially safer than other methods, for both the piglet and the farmer. 3) Emphasize that the timing of EpiFix™ administration aligns with when piglets receive various supplements and vaccines, allowing for ease of adoption. 4) Emphasize that despite EpiFix™ containing hormones, the amount of hormones in meat from EpiFix™-treated pigs is actually lower than in intact pigs due to underdeveloped testes. 5) Highlight the economic value of animals that die following surgical castration and the annual losses incurred by pork producers as a result, while stressing the potential of our technology to mitigate this loss. Changes/Problems:Product name:In our SBIR Phase I proposal, we initially introduced our product as iNeuter™. As our research expanded beyond pigs to other species, particularly companion animals, we realized that the name iNeuter™ is better suited for a male sterilization product in dogs and cats, aligning with the common terminology used for desexing these pets. In the context of livestock, the term "neuter" isn't commonly used to describe male castration. As such, during SBIR Phase I, the product name was changed fromiNeuter™ toEpiFix™, so EpiFix™ will be used in the final report and subsequent Phase II study protocol. The trademark forEpiFix is currently under review by the USPTO. Product design:The Phase I study was originally proposed for the delivery ofiNeuter chemicals (estrogen) in microspheres. However, a long-term preliminary study using estrogenalone in microspheres yielded inconclusive results, suggesting that the inhibition of testis development may not be sustained until slaughter. Later, we discovered that neonatal administration of androgen potentially inhibits both pituitary function and testis development. Therefore, we conducted experiments with a combination of estrogen and androgen to target the hypothalamus, pituitary, and testis simultaneously, aiming to maximize treatment effectiveness in reducing boar taint. Therefore, in the Phase I studies, we hypothesized that a combined treatment of estrogenwith androgencould provide longer-term testis development inhibition and boar taint reduction. Instead of producing estrogen+androgen microspheres,we tested commercially available implants (from Zoetis, Merck, and Elanco) due to regulatory issues associated with using androgens and microspheres. However, we obtained a DEA license for controlled substance schedule III (anabolic steroids) use and developedpellet-typeEpiFix™prototypes through a third-partydrug formulation company.Recently, we secureda patentfor the use of estrogen+androgenimplantsfor the chemical castration of boars (Non-Surgical Prevention of Boar Taint and Aggressive Behavior, US, patent number pending). What opportunities for training and professional development has the project provided?During Phase I, we collaborated with a research laboratory affiliated with the Department of Veterinary Medicine at the University of Illinois at Urbana-Champaign (UIUC). Through a series of small-scale seminars, we provided participating students with theoretical knowledge about swine reproductive organ development and function, and the mechanism responsible for boar taint appearance. We also educated the students on the principles of EpiFix™, which can be applied to other mammalian species and be useful for other academic pursuits. These students had the opportunity to learn histological and molecular biological experiments with samples we provided. We also collaborated with the Meat Science Laboratory in the Animal Science Department of UIUC. The participating students received training in evaluating the quality of pork meat and presence of boar taint. They also had the opportunity to directly practice these skills by using our samples. Recently, we have recruited intern students from UIUC to work on the development of an implant injector specifically designed for pigs. Together, we are in the process of creating a prototype using 3D printing technology. Through this project, we are providing knowledge on scientific concepts related to the product and the opportunity to acquire manufacturing experience. How have the results been disseminated to communities of interest?We are actively collaborating with the Alliance for Contraception in Cats & Dogs (ACC&D), an organization committed to finding non-surgical sterilization treatments for companion animals. We were invited to present our project at their Council of Stakeholders' Annual Briefing held in Tampa (FL) on March 15, 2023. At this meeting, we described the use of EpiFix™ for neutering dogs and cats, as well as its application to reduce boar taint in pigs. Veterinarians affiliated with the ACC&D and representative employees of related pharmaceutical companies attended this meeting, and we received positive responses regarding the potential marketability of EpiFix™. Recently, we had a meeting with Gary Evans, the new president of ACC&D, on January 12, 2024. ACC&D plans to expand its outreach efforts toward different audiences and include information about Insigna's project (https://www.acc-d.org/epivara). Gary also reiterated the interest of ACC&D in our technology, which they consider one of the most promising solutions to replace surgical sterilizations. We had an interview with Smile Politely, a local general interest magazine in Champaign-Urbana (IL), where we discussed the need for alternative sterilization methods like EpiFix™ in the swine industry. Title: "Epivara Inc. has created a single-injection alternative to spays and neutering" (https://www.smilepolitely.com/culture/epivara_inc._has_created_a_single-injection_alternative_to_spays_and_neuter/) Since 2020, we have participated at the annual AgTech Summit (https://researchpark.illinois.edu/all-events/agtechsummit/), organized by the University of Illinois to showcase promising technology solutions that address challenges facing agriculture. Through this platform, either as presenters or with a booth, we have been promoting the functionality and potential marketability of EpiFix™. In January of 2024, we published an article in the online science magazine Research Outreach, which covers the use of our technology in companion animals (https://researchoutreach.org/articles/non-surgical-sterilisation-ispay-pet-spaying-operations/). Although this article did not discuss our swine technology specifically, it was a great opportunity to reach a broad audience who otherwise would be unaware of technological advancements such as ours. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impact Statement:For centuries, physical castration has been the norm on swine farms for managing male pig behavior, fertility, and eliminating boar taint in pork. However, this practice often lacks proper sterilization and pain management, resulting in increased animal suffering, worker morale issues, and animal health risks. Rising concerns about animal welfare are driving a shift away from such methods without anesthesia. In response, Insigna is developing EpiFix™, a single-injection solution designed to replace physical castration in pigs, offering the same benefits without the pain or risks. This USDA SBIR Phase I aimed to validate EpiFix's potential by conducting studies on male pigs, including identifying safe doses, comparing indicators of meat quality, and assessing its impact on testicular development and the reproductive system. Results showed that EpiFix, at the right dosage, successfully achieved the same outcomes as castration. The US swine industry is highly volatile in terms of profits, largely due to increasing production costs, which have risen by 53% since 2020. This has forced farmers to seek cost-effective solutions. Our Phase I research indicates that EpiFix is a viable substitute for surgical castration, offering key benefits like boar taint prevention. Consequently, EpiFix has the potential to reduce piglet morbidity linked with physical castration, thus boosting production yields, financial stability for swine farmers, and promoting animal welfare. The research results according to each objective below are all statistically significant (One-way ANOVA/Student's t-test;P<0.05). Objective (1) identify effective and safe doses of EpiFix that will inhibit testosterone/androstenone production and boar taint in male pigs until slaughtering age (26 weeks of age) For this objective, three main studies were conducted. In all studies, implants with estrogen and androgen were administered to newborn pigs. 1) We identified a promising implant that significantly inhibited or reduced testis development, reducing testes weights by 84%, testosterone (T) levels by 61%, and LH levels by 97% compared to intact boars at 4 weeks after birth, respectively. 2) Implant PK revealed rapid drug release within a month, with complete elimination from the body in 3 months. At slaughter, EpiFix also significantly lowered LH levels in a dose-dependent manner, reducing testes weight by about 82% compared to intact boars. Histological analysis of the testes indicated more underdeveloped Leydig cells, causing a 58% drop in T production. Subsequent studies showed EpiFix-treated pigs had 75% fewer Sertoli cells. Regarding the safety of EpiFix at all tested doses, there were no differences in major organ weights (liver, kidney, spleen) at slaughter compared to the intact group. Serum chemistry analysis showed no differences in biochemical indicators, and the complete blood count test revealed no blood composition changes due to EpiFix treatment. Overall, these results indicate that EpiFix treatment during the neonatal period has no adverse effects on major organs, blood composition, or inflammation in male pigs. 3) EpiFix effectively inhibited testis development until slaughter age, with consistent results regarding serum T/LH, terminal testes weight, and fat androstenone/T. After slaughter, trained panelists gave EpiFix-treated pigs about 70% lower boar taint scores than intact boars. Of note, the mean score of the EpiFix group was even 50% lower than barrows. These studies support EpiFix as a non-surgical alternative to castration, effectively limiting testes function by inhibiting Leydig and Sertoli cells. We also identified the importance of a temporary LH increase before 4 weeks of age for testicular development. These results were used to apply for a patent, which was allowed on December 6th, 2023 (not yet assigned). More data will be shared in a future peer-reviewed publication. Throughout these studies, we have acquired and validated valuable chemical analysis tools for future research and product optimization. Through testing different EpiFix prototypes and drug doses, we discovered a safe and effective implant and revealed its PK. We are currently utilizing this novel data to develop an optimized prototype through a third-party contractor. We initiated the regulatory approval process for EpiFix by opening an Investigation New Animal Drug (INAD) file with the FDA. The insights gained from this initial testing will be crucial in designing rigorous pivotal studies required for drug approval. Objective (2) compare the circulation/tissue level of androstenone and skatole (boar taint indicators) and meat quality in EpiFix-treated, castrated, and intact boars For this objective, one study was conducted in which EpiFix was administered to newborn pigs one day after birth and its safety and effectiveness was evaluated until 24 weeks of age. The evaluations included boar taint indicators, meat quality, and analysis of residual amounts of drugs in meat post-slaughter (consumer safety). In the study, fat androstenone, linked to boar taint, was 72% lower in treated pigs compared to intact boars. Although EpiFix pigs had slightly lower body weights at slaughter, meat quality assessments revealed desirable thinner back fat, a key indicator of carcass yield, compared to intact boars. No differences were found in other meat quality factors like hot carcass weight, loin eye area, dressing percentage, marbling, or firmness compared to the control group. Residue analysis in meat of 24-week-old pigs showed drug levels below detection, ensuring consumer safety. Although quantitative analysis of skatole was not performed in this study, the results presented were sufficient to conclude that EpiFix prevents boar taint. Industry experts indicated the potential impact on EpiFix marketability due to carcass weight's significant role in profit. Our future focus (Phase II) will be on optimizing EpiFix dosage to preserve benefits without affecting growth. Objective (3) determine if EpiFix inhibits functional development of the testis and Kisspeptin expression in the hypothalamus For this objective, one study was conducted in which newborn pigs were administered EpiFix and then raised for 8 weeks to evaluate the treatment's effectiveness in inhibiting testes development and reducingKISS1expression. At 8 weeks old, all individuals were sacrificed and their testis and hypothalamic tissues were collected for analysis. Histological analysis of the treated pigs' testes showed Leydig cells remained in the undifferentiated neonatal stage. Both serological and histological data from prior studies indicated that this resulted in low concentration of male hormones until the time of slaughter. QuantifyingKISS1in hypothalamic tissues revealed a 73% reduction with EpiFix compared to intact boars. This indicates that EpiFix can diminish gonadal function in male pigs, reducing boar taint by suppressing Kisspeptin neuron development. These findings mark a significant milestone for Insigna, confirming that neonatal estrogen and androgen treatment disrupts kisspeptin system maturation. This novel concept, not previously documented, offers promise as an alternative to surgical castration. This data suggests that the kisspeptin system is well-conserved across mammals, indicating the potential to expand the applicability of EpiFix to other species.

Publications

Progress 07/01/22 to 02/28/23

Outputs
Target Audience:Perception and Use of Epivara's Product (iNeuter)in Swine Production Introduction The National Pork Producers Council reports that over 60,000 pork producers in the US generate annual revenues of over $20 billion. Additional sectors tied to pig production, including transportation, animal care, and consumer-related purchases, bring the entire industry to an estimated $122 billion. Our company's target market is aimed at two substantial aspects of this industry: the castration of male pigs and the grow-out business. According to USDA data from 2020, the US produced more than 78 million pigs, which equates to over 10% of the global supply. Assuming 50% of the pigs taken to market are male, our target opportunity stands at just under 40 million pig castrations per year in the US alone. Initially, our focus will be the US pork producers, before we expand to producers globally. To better understand the industry and its needs, we've conducted several conversations with swine veterinarians and pork producers, exploring the viability of our new product, iNeuter. Our effort is to position iNeuter as an alternative to surgical castration or Improvest, the only FDA-approved product currently being used to delay the onset of puberty in male pigs. Insights from a Swine Veterinarian The veterinarian highlighted the following points: Improvest offers performance benefits such as improved feed conversion rate (FCR) and growth rate by allowing pigs to remain boars for a longer period, up until 3-5 weeks before slaughter. Our new product, iNeuter, may not offer these same performance benefits. Resistance to adopting the use of Improvest comes primarily from retailers due to concerns associated with genetically modified organisms (GMO). Despite the resistance, the benefits of Improvest, including a net gain of $4-5 per pig after accounting for the cost of administration, outweigh the inconveniences of administering two doses. Labor and safety costs associated with castration are minimal, with occasional injuries from blade use not being a significant concern. Views from a Large-Scale Hog Producer The producer discussed several industry practices, public perceptions, and the potential of iNeuter: The producer expressed willingness to try iNeuter, though they believe customer acceptance is unlikely. Castration typically occurs within 3-5 days of birth in the industry. Improvest has high brand awareness and offers significant production advantages, estimated at $2.5-$4.5 more per pig. The producer sees a single early shot as a major management advantage, especially since it would be given at the traditional time reserved for castration. Public perception of Improvest is mixed. Despite evidence of its benefits, the public does not seem to perceive these advantages. Concerns exist about the human safety of handling iNeuter, similar to those associated with Improvest. The producer's market currently does not use Improvest due to customer policies, notably Clemens Foods. Animal welfare and hormone-related concerns are weighed against each other, with hormone issues often considered more important. Castration costs range from $0.15-$0.20 per pig, but poor technique or genetic issues can result in significant losses, up to 50-75 pigs per week in a farm that weans 5,000 pigs per week. The producer is interested in the human safety, administration, and potential residue of iNeuter compared to Improvest, particularly at 4 months. Market Research Insights Our research on various national markets concluded that targeting the Brazilian market would likely be easier than focusing on hormone-sensitive markets such as South Korea or Europe. Conclusion Understanding the views of industry experts and large-scale producers is critical for the potential adoption and success of new products like iNeuter. While concerns exist, there is a recognition of the benefits and potential improvements these alternatives can bring to swine production. However, customer acceptance and market dynamics will ultimately play a significant role in their widespread adoption. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this study, we collaborated with a research laboratory affiliated with the Department of Veterinary Medicine at the University of Illinois at Urbana-Champaign (UIUC). Through a series of small-scale seminars, we provided participating students with theoretical knowledge about swine reproductive organ development and function, and the mechanism of occurrence of boar taint. We also educated the students on the principles of iNeuter that can be applied in other academic pursuits. These students had the opportunity to learn histological and molecular biological experiments using our samples. We also collaborated with the Meat Science Laboratory in the Animal Science Department of UIUC. The participating students received training in evaluating the quality of meat and boar taint, and had the opportunity to directly participate in experiments using our samples. How have the results been disseminated to communities of interest?We participated as presenters at the ACC&D Council of Stakeholders' Annual Briefing held in Tampa, Florida, on March 15, 2023. At this meeting, we explained the use of iNeuter for neutering dogs and cats, as well as technology to reduce boar taint in pigs. Veterinarians affiliated with ACC&D and representative employees of related pharmaceutical companies attended this meeting, and we received positive responses regarding the potential marketability of iNeuter. (https://www.acc-d.org/sex-steroids) We had an interview with Smile Politely, where we discussed the need for alternative sterilization methods like iNeuter in the swine industry. Title: "Epivara Inc. has created a single-injection alternative to spays and neutering" (https://www.smilepolitely.com/culture/epivara_inc._has_created_a_single-injection_alternative_to_spays_and_neuter/) Since 2020, we have been participating annually in the Agtech Summit (https://researchpark.illinois.edu/all-events/agtechsummit/), either as presenters or with a booth. Through this platform, we have been promoting the functionality and potential marketability of iNeuter. What do you plan to do during the next reporting period to accomplish the goals?During Phase I, we were able to measure boar taint from some samples and verify the effectiveness of iNeuter, but there are still samples that have not yet been analyzed. By analyzing these, we anticipate being able to confirm the more definitive boar taint reduction effect of iNeuter. Therefore, during the remaining research period, we will accurately measure boar taint from the secured samples. This involves trained and validated individuals directly smelling the samples, so we are currently preparing an Institutional Review Board (IRB) for this task. Based on the data obtained during Phase I, we will file for a US/PCT patent for technology to reduce boar taint through the inhibition of male pig reproductive organ development using iNeuter. Through interactions with swine industry workers, we will listen to market demands and establish a Phase II research plan that reflects these requirements. In particular, we will conduct surveys on the ease of use in the field and concerns about iNeuter. In preparation for the more advanced safety/functionality research on iNeuter to be conducted during Phase II, we will interact with relevant researchers to prepare large-scale feed-efficiency and behavior tests.

Impacts
What was accomplished under these goals? The surgical castration of pigs can be a painful and stressful procedure for the animals, as it is done without anesthetics or analgesics. This can also pose risks to farm workers. As one may imagine, castration of pigs is one of the least desirable jobs in a swine operation. iNeuter™ has the potential to significantly improve animal welfare and employee morale in this industry. iNeuter is a non-surgical method of castration that uses a single injection of APIs. This prevents the development of the testes and the production of androgens, which are responsible for boar taint. iNeuter has several advantages over other methods of castration. iNeuter is a non-surgical method that is safer and easier to perform than surgical castration. The injection can be incorporated into the existing schedule of vaccine and iron injections in neonates. This could not only reduce the cost of treatment and risk of injury for farm workers, but also lead to improved employee morale and reduced turnover. iNeuter is a proactive response to the growing public interest in animal welfare. There has been a surge of EU regulations associated with the castration procedure, and a number of countries either have banned or proposed banning surgical castration of pigs. iNeuter technology could proactively address such animal welfare concerns, getting in front of an animal welfare and public sentiment outcry that is making its way to the US. The USDA SBIR Phase I study aimed to evaluate the capability of iNeuter, administered via a single neonatal injection, to replace surgical castration in newborn male pigs. The treatment is anticipated to inhibit testicular development, reduce androgen production, prevent aggressive behavior, and decrease boar taint in meat. The objectives included assessing iNeuter's effect on hypothalamic KISS1 expression, identifying effective and safe doses that suppress testosterone and reduce boar taint, and comparing iNeuter with physical castration and immunocastration methods. The experiments employed a microsphere form of iNeuter and a similar injectable implant used for beef cattle. Pigs were treated with iNeuter immediately post-birth, and evaluations were conducted before 26 weeks of age. Based on active ingredient quantity, iNeuter implants were categorized into low, middle, and high-dose groups. The findings suggested that medium to high iNeuter doses effectively inhibited testicular development and reduced androgen levels and boar taint up to slaughter age. 1) Major activities completed/experiments conducted: Study 1- Various combinations of drug implants were administered to newborn pigs. Study 2- The efficacy and safety of low-dose iNeuter and pharmacokinetics of the chemicals/Drugs deemed effective in inhibiting testis development were selected through Study 1. These drugs were then incorporated into low-dose implants, which were implanted into newborn pigs. Study 3- Efficacy and safety of high-dose iNeuter: In comparison to Study-2, implants with a relatively higher dose were implanted into newborn pigs. Study 4- Comparative study of commercial implants: Commercial implants of various doses were administered to newborn pigs, and their safety and effectiveness were analyzed. 2) Data collected First, we observed the drug release patterns of various drug implants in order to incorporate them into the design of future prototypes. We also compared and analyzed drug release patterns with the final outcomes from our efficacy and safety studies. Efficacy: 1) measuring androgen levels in the blood; 2) measuring levels of LH hormone in the blood; 3) quantitative analysis of boar taint-causing molecules; 4) boar taint inspection by a human panel; 5) comparison of KISS1 gene expression in the hypothalamus; 6) analysis of meat quality; and 7) comparison of growth rates. Safety: 1) Complete Blood Counting; 2) Serum Chemistry; 3) comparison of the weight of major organs; and 4) testing for residual drug amounts in meat and blood. 3) Summary statistics and discussion of results Efficacy analysis showed that the drug was effectively delivered within two months of treatment. The active ingredient in iNeuter significantly lowered the level of Luteinizing Hormone (LH), which is crucial for initial testicular development, leading to smaller testes compared to the untreated group. This was attributed to iNeuter hindering the maturation of Leydig cells in the testes, leading to reduced male hormone concentration till the time of slaughter. Furthermore, iNeuter decreased the level of androstenone, a compound responsible for boar taint. Although boar taint scores were lower in iNeuter-treated pigs compared to the control group, they were higher than physically castrated barrows, suggesting that iNeuter's performance requires further optimization. iNeuter was found to suppress the expression of the KISS1 gene in the hypothalamus, a key player in the reproductive system. This ability of iNeuter to stifle the development of the Kisspeptin system, coupled with its effects on pituitary and testicular somatic cells, results in loss of gonadal function in male pigs, further reducing boar taint. Assessment of meat quality showed that iNeuter-treated pigs had less back fat than the control group, leading to a higher muscle ratio in the loin meat of the same weight. However, other meat quality parameters remained unchanged. While iNeuter's efficacy in reducing boar taint was not as high as its competitor Improvest, further research optimizing the dose and exposure length could improve its performance. The safety evaluation of iNeuter showed no adverse effects on the development and function of major organs like liver, kidney, and spleen. In addition, no significant changes in blood cell composition or inflammation markers were observed following iNeuter treatment. At slaughter, pigs from the low-middle-dose group had similar weights to the control group and the physically castrated barrows, while the high-dose group had lower weights, suggesting dosage consideration for future product development. Drug residue tests conducted post-treatment indicated that drug levels in meat and blood were below detectable levels, ensuring that consumers would not be exposed to these drugs. 4) Key outcomes or other accomplishments realized Proof of concept: Through experimentation, it has been proven that iNeuter is able to suppress the development and function of the reproductive system by inhibiting the development of the Kisspeptin neuron system. This inhibitory effect of iNeuter on testicular development ultimately reduced the boar taint in male pigs without the need for physical castration. The effective dose of iNeuter: Through Phase I research, we have secured a safe dose of iNeuter with active properties and have understood its pharmacokinetics. Based on these results, we are currently preparing for prototype production through a third-party contractor. FDA regulation-INAD: Based on the results we've obtained thus far, we are preparing to open an INAD (Investigational New Animal Drug) and have currently submitted a Sponsor Fee Waiver request to minimize the cost of the FDA approval process. We are aiming to open the INAD in August-September of 2023. Scientific findings: The study revealed that iNeuter can effectively inhibit testicular function in pigs, offering an alternative to physical castration. It acts on the Kisspeptin system, conserved across mammals, indicating potential for wider applicability. iNeuter, directly and indirectly, curbs the development of Leydig and Sertoli cells, pivotal to testicular maturation. The study identified a key role for Luteinizing Hormone (LH) in postnatal testicular development. These findings led to a patent application and will be detailed in a forthcoming peer-reviewed paper. The study also bolstered chemical analysis tools for future product development.

Publications